期刊
CRYOBIOLOGY
卷 95, 期 -, 页码 151-156出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.cryobiol.2020.03.009
关键词
Oogonia; Freezing; Vitrification; Hypothermic storage; Transplantation; Surrogate production
资金
- National Research Development and Innovation Office of Hungary [FK 124585]
- Fisheries Fund for Renewable Fisheries by the EU and Hungary
- Stipendium Hungaricum Scholarship Programme [106360, 192128, 218533]
- EU [EFOP-3.6.3-VEKOP-16-2017-00008]
- European Social Fund
- Ministry of Human Capacities within the framework of water related research of Szent Istvan University [1783-3/2018/FEKUTSRAT]
The aim of this study was to develop short- and long-term preservation protocols for European eel ovarian stem cells (OSCs) through hypothermic storage and cryopreservation of ovarian fragments that will assist in current conservation programs of this critically endangered species. Firstly, a freezing procedure was developed by testing different cryomedia and technical aspects of freezing. Utilization of 1.5 M of dimethyl sulfoxide (Me2SO), 0.1 M glucose and 1.5% BSA yielded optimal OSCs survival. Additionally, equilibration of 50-mg ovarian fragments for 30 min and plunging into lN(2) at -80 degrees C displayed the highest OSC viability. Different cooling rates ranging from -1 to -40 degrees C/min did not significantly affect OSC viability when thawing in a 10 degrees C water bath. In addition, application of needle-immersed vitrification (NIV), combining ES3 (1.5 M PG and 1.5 M Me2SO) with VS3 (3 M PG and 3 M Me2SO) yielded the highest viability rates. Finally, hypothermic storage (4 degrees C) of ovarian fragments and ovarian cell suspensions displayed favorable viability of similar to 90% after 48 h of storage and similar to 65% after 72 h of storage. The development of OSC preservation methods presents an onset of further development of germline stem cell (GSC) manipulation techniques in this species. Cryopreservation of OSCs can enable a continuous supply of cells for either transplantation or in vitro cell culture thus enabling new and improved management and conservation strategies for this endangered species.
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